pMD18-T+Vector圖譜

1、pMD18-T VectorD101A1 pg500pMD19-T VectorD102A1 pg50031 BIO VIEW No2# BIO VIEW No2pMD18-T Vector pMD19-T Vector pUC18 pUC19PCR(TA Cloning) <ba I Sal I DNAPCREcoR VPCRpUC18 pUC19 EcoRV 3*3T"A"PCRpUC18 pUC19pUC18pUC19Ligation Mix(5 )Control Insert (500 bp)ControlpMD19-T VectorpMD18-T Vecto

2、rpMD19-T VectorP-PMD18-T & pMD19-T VectorpMD18-T VectorflcaBEST1M Sequencing Primer RV-MGAGCGGATAACAATTTCACACAGGXmaiEcoRVCloning SiteHine 11Acc I Sse6387 ISph 1 Hind 111Kpn； Xbai EcoRV| PstlGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGATATCGTCGACCTGCAGGCATGCAAG

3、CTTlcZEccR l ScIGGCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGCAGCACTGACCCTTTTGGGACCGCBcaBEST Sequencing Pnmer M13-47Digested by EcoR VpMD19-T Vector let agaga t T a t eg t cgacc - .-agatet c ta Ttagcagc tgg -(T CIorwg Site)BcaBEST Sequmcing Primer RV MGAGCGGATAACAATTTCACACAGGHine ll S$e8387 I Acc

4、jXma I Hind III Sp/H |Pstl I Sall EcoRV Xbf I 嚴Saci fcoR IGAGCGGATAACAATTTCACACAGG AAACAGCTATGACCATGATTACGCCAAGCTTGCATGCCTGCAGGTCGACGATATCTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGCAGCACTGACCCTTTTGGGACCGCBcaBEST" Sequencing Primer Ml3-47gtcgacgatT caget gc taD

5、igested by fooR Vatetetaga.Ttagagatct. ("lomnghe)pMD18-T orpMD19-T Vector (50 ng/pl)Control Insert (50 ng/pl)10 pl 130 pl 5Ligation Mix*# BIO VIEW No2# BIO VIEW No2B.# BIO VIEW No2# BIO VIEW No2ControlInsert(efu/pg Vector)(%)(%)*L W2.8 10°96.3902.2 10、40.20# BIO VIEW No2# BIO VIEW No2Bea#

6、BIO VIEW No2# BIO VIEW No2Cloning site419BcaBEST Sequencing Primer M13-47 binding site 347 370BcaBEST Sequencing Primer RV-M binding site478 500LacZ operator146 469ColE1 ori852* 1466Amp1626-2486Cloning site431BcaBEST Sequencing Primer M13-47 binding site 347 370DcaD匚ST Sequencing Primer RV-M binding site478 - 500LacZ operator146 469ColE1 ori8521466Amp1626 2486A.PpMD1 &T or pMD19-T Vector*1Control Insert*2dH2O3glpMD18-T or pMD19-T Vector* 11Insert DNA*30.1 pmol 0.3 pmoldHzOup to 5 pl33 BIO VIEW No2LacZLac